The Molecular Regulative Functions Of The Arabidopsis Transcription Factor TDF1 In Anther Development

In Arabidopsis, TDF1 encodes a MYB transcription factor which plays a vital role in tapetal development and function during anther development. TDF1 was highly expressed in meiocytes, tapetum and microspores in anther development.The tdfl mutant shows a complete male sterile phenotype. In this study, the TDF1 gene was obtained by PCR method and cloned into the expression vector pET-32a. After the prokaryotic expression vector'TDF1-pET32a'introduced into E. coli BL21(DE3)., the TDF1 recombinant protein of 56.2 KD was expressed by induction with IPTG. The fusion protein majorly existed in the inclusion body. After extracted from the inclusion body, the TDF1 protein was used as the antigen to immune rabbits. ELISA assay showed that the titer of the prepared polyclonal antibody was 1:2500. The immunological specificity of the polyclonal antibody to TDF1 was confirmed by Western blot analysis. The prepared antibody in this work should facilitate the further functional investigation of TDF1 in biochemical levels in Arabidopsis anther development. Bioinformatics analysis identified several transcription factors may be involved in the regulation of TDF1. Furthermore, the transgenic line of TDF1 and TAP-tag fusion protein was obtained, which provide easy access to find the direct target of TDF1 by ChIP. These works lay the foundation for further investigation of the molecular basis of TDF1 in anther development.